Li W, Chi K, Gallick G E, Chan J C
Virology. 1987 Jan;156(1):91-100. doi: 10.1016/0042-6822(87)90439-9.
Previously, we reported the monoclonal antibody detection of transformation-associated proteins (TAP) in ts110 murine sarcoma virus-transformed normal rat kidney (6M2) cells (Chan et al., 1986). In this study, we used the same monoclonal antibody to investigate the subcellular localization, the fate and the mitogenic activity of TAP, as well as the correlationship between TAP synthesis and the expression of transformation properties of 6M2 cells. It was found that TAP were localized in the cytoplasm (probably the Golgi apparatus) of 6M2 cells. TAP were found as three intracellular polypeptides (mol wt of 66K, 63K, and 60K, respectively), and were rapidly released into extracellular medium. Upon release, TAP changed to two extracellular polypeptides (mol wt of 68K and 64K, respectively). Furthermore, the synthesis of TAP was temperature sensitive and correlated closely with the expression of transformation properties of the 6M2 cells. TAP have been purified by monoclonal antibody-affinity column chromatography and found to have a synergistic effect with insulin in stimulating the DNA synthesis of normal rat kidney cells.
此前，我们报道了用单克隆抗体检测ts110鼠肉瘤病毒转化的正常大鼠肾（6M2）细胞中的转化相关蛋白（TAP）（Chan等人，1986年）。在本研究中，我们使用相同的单克隆抗体来研究TAP的亚细胞定位、命运和促有丝分裂活性，以及TAP合成与6M2细胞转化特性表达之间的相关性。结果发现，TAP定位于6M2细胞的细胞质中（可能是高尔基体）。TAP以三种细胞内多肽形式存在（分子量分别为66K、63K和60K），并迅速释放到细胞外培养基中。释放后，TAP转变为两种细胞外多肽（分子量分别为68K和64K）。此外，TAP的合成对温度敏感，并且与6M2细胞转化特性的表达密切相关。TAP已通过单克隆抗体亲和柱层析纯化，并发现其在刺激正常大鼠肾细胞的DNA合成方面与胰岛素具有协同作用。
